The biochemical effects of extracellular Zn2+and other metal ions are severely affected by their speciation in cell culture media

Metallomics ◽  
2015 ◽  
Vol 7 (1) ◽  
pp. 102-111 ◽  
Author(s):  
H. Haase ◽  
S. Hebel ◽  
G. Engelhardt ◽  
L. Rink
Keyword(s):  
Cell Culture ◽  
Bovine Serum Albumin ◽  
Culture Medium ◽  
Culture Media ◽  
Zinc Homeostasis ◽  
Cell Culture Medium ◽  
Cell Culture Media ◽  
Biochemical Effects ◽  
Cellular Environment

Differential speciation and lower zinc buffering by less bovine serum albumin (BSA) in cell culture medium lead to altered zinc homeostasis compared to the cellular environmentin vivo.

scholarly journals Selectivity of direct plasma treatment and plasma-conditioned media in bone cancer cell lines

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Inès Hamouda ◽  
Cédric Labay ◽  
Uroš Cvelbar ◽  
Maria-Pau Ginebra ◽  
Cristina Canal
Keyword(s):  
Cell Culture ◽  
Plasma Treatment ◽  
Cancer Cell ◽  
Culture Medium ◽  
Culture Media ◽  
Bone Cancer ◽  
Reactive Species ◽  
Conditioned Media ◽  
Cell Culture Medium ◽  
Cell Culture Media

AbstractAtmospheric pressure plasma jets have been shown to impact several cancer cell lines, both in vitro and in vivo. These effects are based on the biochemistry of the reactive oxygen and nitrogen species generated by plasmas in physiological liquids, referred to as plasma-conditioned liquids. Plasma-conditioned media are efficient in the generation of reactive species, inducing selective cancer cell death. However, the concentration of reactive species generated by plasma in the cell culture media of different cell types can be highly variable, complicating the ability to draw precise conclusions due to the differential sensitivity of different cells to reactive species. Here, we compared the effects of direct and indirect plasma treatment on non-malignant bone cells (hOBs and hMSCs) and bone cancer cells (SaOs-2s and MG63s) by treating the cells directly or exposing them to previously treated cell culture medium. Biological effects were correlated with the concentrations of reactive species generated in the liquid. A linear increase in reactive species in the cell culture medium was observed with increased plasma treatment time independent of the volume treated. Values up to 700 µM for H2O2 and 140 µM of NO2− were attained in 2 mL after 15 min of plasma treatment in AdvDMEM cell culture media. Selectivity towards bone cancer cells was observed after both direct and indirect plasma treatments, leading to a decrease in bone cancer cell viability at 72 h to 30% for the longest plasma treatment times while maintaining the survival of non-malignant cells. Therefore, plasma-conditioned media may represent the basis for a potentially novel non-invasive technique for bone cancer therapy.

scholarly journals Serum-free medium increases the replication rate of the avian coronavirus infectious bronchitis virus in chicken embryo kidney cells

2021 ◽  
Author(s):  
José A. Quinteros ◽  
Glenn F. Browning ◽  
Amir H. Noormohammadi ◽  
Mark A. Stevenson ◽  
Mauricio J. C. Coppo ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Cultures ◽  
Infectious Bronchitis Virus ◽  
Chicken Embryo ◽  
Culture Medium ◽  
Culture Media ◽  
Cell Culture Medium ◽  
Cell Culture Media ◽  
Infectious Bronchitis ◽  
Porcine Epidemic Diarrhoea Virus

AbstractInfectious bronchitis virus (IBV), an avian coronavirus, can be isolated and cultured in tracheal organ cultures (TOCs), embryonated eggs and cell cultures. TOCs and embryonated eggs are commonly used for viral isolation but use of these is laborious and expensive. Cell cultures have been used only with IBV strains that have previously been adapted to grow under laboratory conditions, and not for primary isolation. Previous studies using the coronavirus porcine epidemic diarrhoea virus (PEDV) have suggested that foetal bovine serum (FBS), a common component of cell culture media, can inhibit the adsorption of coronaviruses onto the host cell membrane receptors. In the present study, the replication of IBV in primary chicken embryo kidney (CEK) cell cultures and the Leghorn hepatocellular carcinoma (LMH) cell line was examined using two different cell culture media, one containing FBS and the other containing yeast extract (YE). A reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) assay was used to quantify viral RNA copies in cell lysates. The highest concentrations of viral genomes were observed when the cell culture medium did not contain FBS. Examination of the infectivity of virus grown in CEK cell cultures was examined by titration in embryonated chicken eggs, demonstrating that the cell lysate from CEK cell cultures in medium without FBS contained a higher median embryo infectious dose (EID50) than that from CEK cell cultures in medium containing FBS. These results suggest that improved replication of IBV in cell cultures can be achieved by the omission of FBS from the cell culture medium. This may enhance the potential for production of vaccines in cell culture and facilitate the isolation of emergent IBV strains in cell cultures.

scholarly journals The Cell Culture Medium Affects Growth, Phenotype Expression and the Response to Selenium Cytotoxicity in A549 and HepG2 Cells

Antioxidants ◽  
2019 ◽  
Vol 8 (5) ◽  
pp. 130 ◽  
Author(s):  
Lisa Arodin Selenius ◽  
Marita Wallenberg Lundgren ◽  
Rim Jawad ◽  
Olof Danielsson ◽  
Mikael Björnstedt
Keyword(s):  
Cell Culture ◽  
Cell Growth ◽  
Culture Medium ◽  
Culture Media ◽  
A549 Cells ◽  
Culture Conditions ◽  
Cell Culture Medium ◽  
Cell Culture Media ◽  
Toxic Response ◽  
Selenium Compounds

Selenium compounds influence cell growth and are highly interesting candidate compounds for cancer chemotherapy. Over decades an extensive number of publications have reported highly efficient growth inhibitory effects with a number of suggested mechanisms f especially for redox-active selenium compounds. However, the studies are difficult to compare due to a high degree of variations in half-maximal inhibitor concentration (IC50) dependent on cultivation conditions and methods to assess cell viability. Among other factors, the variability in culture conditions may affect the experimental outcome. To address this, we have compared the maintenance effects of four commonly used cell culture media on two cell lines, A549 and HepG2, evaluated by the toxic response to selenite and seleno-methylselenocysteine, cell growth and redox homeostasis. We found that the composition of the cell culture media greatly affected cell growth and sensitivity to selenium cytotoxicity. We also provided evidence for change of phenotype in A549 cells when maintained under different culture conditions, demonstrated by changes in cytokeratin 18 (CK18) and vimentin expression. In conclusion, our results have shown the importance of defining the cell culture medium used when comparing results from different studies.

scholarly journals Extracellular electrical recording of pH-triggered bursts in C6 glioma cell populations

2016 ◽  
Vol 2 (12) ◽  
pp. e1600516 ◽  
Author(s):  
Paulo R. F. Rocha ◽  
Maria C. R. Medeiros ◽  
Ulrike Kintzel ◽  
Johannes Vogt ◽  
Inês M. Araújo ◽  
...  
Keyword(s):  
Cell Culture ◽  
Ion Channels ◽  
Electrical Activity ◽  
Glioma Cell ◽  
Culture Medium ◽  
Current Noise ◽  
Cell Culture Medium ◽  
Large Area ◽  
Brain Physiology

Glioma patients often suffer from epileptic seizures because of the tumor’s impact on the brain physiology. Using the rat glioma cell line C6 as a model system, we performed long-term live recordings of the electrical activity of glioma populations in an ultrasensitive detection method. The transducer exploits large-area electrodes that maximize double-layer capacitance, thus increasing the sensitivity. This strategy allowed us to record glioma electrical activity. We show that although glioma cells are nonelectrogenic, they display a remarkable electrical burst activity in time. The low-frequency current noise after cell adhesion is dominated by the flow of Na+ions through voltage-gated ion channels. However, after an incubation period of many hours, the current noise markedly increased. This electric bursting phenomenon was not associated with apoptosis because the cells were viable and proliferative during the period of increased electric activity. We detected a rapid cell culture medium acidification accompanying this event. By using specific inhibitors, we showed that the electrical bursting activity was prompted by extracellular pH changes, which enhanced Na+ion flux through the psalmotoxin 1–sensitive acid-sensing ion channels. Our model of pH-triggered bursting was unambiguously supported by deliberate, external acidification of the cell culture medium. This unexpected, acidosis-driven electrical activity is likely to directly perturb, in vivo, the functionality of the healthy neuronal network in the vicinity of the tumor bulk and may contribute to seizures in glioma patients.

Using 2H2O to study the influence of feeding on protein synthesis: effect of isotope equilibration in vivo vs. in cell culture

2005 ◽  
Vol 288 (6) ◽  
pp. E1277-E1283 ◽  
Author(s):  
Danielle A. Dufner ◽  
Ilya R. Bederman ◽  
Daniel Z. Brunengraber ◽  
Nadia Rachdaoui ◽  
Faramarz Ismail-Beigi ◽  
...  
Keyword(s):  
Cell Culture ◽  
Protein Synthesis ◽  
Steady State ◽  
Culture Medium ◽  
Body Water ◽  
Cell Culture Medium ◽  
Plasma Albumin ◽  
Single Meal

We previously reported that 2H2O can be used to measure rates of protein synthesis during prolonged steady-state conditions (Previs SF, Fatica R, Chandramouli V, Alexander JC, Brunengraber H, and Landau BR. Am J Physiol Endocrinol Metab 286: E665-E672, 2004). The underlying premise of our method is that following the administration of 2H2O, 2H atoms in body water rapidly equilibrate with free alanine before it is incorporated into newly synthesized proteins. We have now directly examined whether 2H2O can be used to measure the influence of a single meal on protein synthesis. In addition, we have compared the use of 2H2O for measuring rates of protein synthesis in vivo vs. in cell culture. Using a rat model, we observed rapid equilibration between 2H in body water and free alanine; therefore we were able to study the response of protein synthesis to a single meal. We observed that ∼50% of the plasma albumin that is synthesized over the course of 24 h is made within ∼5 h after eating (in rats trained to eat a complete 24-h ration of food in a single meal). Contrary to what we observed in vivo, feeding (the replenishment of cell culture medium) does influence the use of 2H2O for in vitro studies. In particular, since there can be slow equilibration of 2H between water and alanine in the cell culture medium, special consideration must be made to avoid underestimating the rate of protein synthesis in vitro.

scholarly journals Anti-Depressive Effectiveness of Baicalin In Vitro and In Vivo

Molecules ◽  
2019 ◽  
Vol 24 (2) ◽  
pp. 326 ◽  
Author(s):  
Li Liu ◽  
Yu Dong ◽  
Xin Shan ◽  
Lin Li ◽  
Baomei Xia ◽  
...  
Keyword(s):  
Cell Culture ◽  
Pc12 Cells ◽  
Culture Medium ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mild Stress ◽  
Cell Culture Medium ◽  
Tnf Α ◽  
Immobility Time

Baicalin (BA), a major polyphenol compound isolated from the extracts of Scutellaria radix, has been previously reported to ameliorate depressive-like behaviors in mice with chronic unpredictable mild stress (CUMS). However, its underlying antidepressant mechanisms remain unclear. This study was designed to confirm the antidepressant-like effects of BA on CUMS induced behavioral abnormalities in mice, and sought to explore the pharmacological mechanisms in vivo and in vitro. The CUMS procedure was carried out to induce depression in mice. Afterwards, the tail suspension test (TST), forced swim test (FST), and open field test (OFT) were performed within 24 h, then sucrose preference test (SPT) was conducted. Additionally, PC12 cells were pretreated with BA for 2 h, then further stimulated with corticosterone for 24 h. The levels of Interleukin-1β (IL-1β), IL-6 and Tumor Necrosis Factor-α (TNF-α) in serum, hippocampus homogenate and cell culture medium were determined using the enzyme-linked immunosorbent assay (ELISA) method. The protein expressions of inhibition of high mobility group box 1 protein (HMGB1)/Toll-like receptor 4 (TLR4)/nuclear factor kappa B (NF-κB) pathways in hippocampus and PC12 cells were detected. Our results showed that CUMS-treated mice presented notable depressive-like symptoms, such as decreased sucrose consumption, increased FST and TST immobility time. While BA (25, 50 mg/kg) significantly attenuated these changes. Besides, BA treatment considerably inhibited inflammatory cytokinesl (IL-1β, IL-6, TNF-α) levels in serum, hippocampus homogenate and cell culture medium. Western blot analysis indicated that BA inhibited the expressions of HMGB1, TLR4, and p-NF-κBp65 both in vivo and in vitro. In conclusion, the present study confirmed that BA possessed efficient antidepressant effects on depression, which was possibly related to the inhibition of HMGB1/TLR4/NF-κB pathways.

Tribological Performance of Cell-Treated Nickel Matrix

2005 ◽  
Author(s):  
Bing Shi ◽  
Thomas Kuhn ◽  
Lawrence Duffy ◽  
Hong Liang
Keyword(s):  
Cell Culture ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Dry Friction ◽  
Culture Media ◽  
Tribological Performance ◽  
Nickel Matrix ◽  
Cell Culture Media ◽  
Pin On Disk

In this study, effects of cell culture on surface properties, and tribological performance were investigated. The wettability of Ni under dry and lubricated conditions, as well as cell-cultured specimens was evaluated. The tribological performance of these samples was compared using a pin-on-disk tribometer. Dry friction tests were conducted and compared with the bovine serum albumin (BSA) solution lubricated Ni and the cell culture media lubricated Ni. The lubrication behavior was discussed and new biofluid mechanisms were proposed.

scholarly journals In Vitro Bioavailability of the Hydrocarbon Fractions of Dimethyl Sulfoxide Extracts of Petroleum Substances

2020 ◽  
Vol 174 (2) ◽  
pp. 168-177 ◽  
Author(s):  
Yu-Syuan Luo ◽  
Kyle C Ferguson ◽  
Ivan Rusyn ◽  
Weihsueh A Chiu
Keyword(s):  
Cell Culture ◽  
Protein Binding ◽  
Aromatic Compounds ◽  
Culture Media ◽  
In Vitro Testing ◽  
Cell Culture Media ◽  
Chemical Profiles ◽  
In Vivo Extrapolation

Abstract Determining the in vitro bioavailable concentration is a critical, yet unmet need to refine in vitro-to-in vivo extrapolation for unknown or variable composition, complex reaction product or biological material (UVCB) substances. UVCBs such as petroleum substances are commonly subjected to dimethyl sulfoxide (DMSO) extraction in order to retrieve the bioactive polycyclic aromatic compound (PAC) portion for in vitro testing. In addition to DMSO extraction, protein binding in cell culture media and dilution can all influence in vitro bioavailable concentrations of aliphatic and aromatic compounds in petroleum substances. However, these in vitro factors have not been fully characterized. In this study, we aimed to fill in these data gaps by characterizing the effects of these processes using both a defined mixture of analytical standards containing aliphatic and aromatic hydrocarbons, as well as 4 refined petroleum products as prototypical examples of UVCBs. Each substance was extracted with DMSO, and the protein binding in cell culture media was measured by using solid-phase microextraction. Semiquantitative analysis for aliphatic and aromatic compounds was achieved via gas chromatography-mass spectrometry. Our results showed that DMSO selectively extracted PACs from test substances, and that chemical profiles of PACs across molecular classes remained consistent after extraction. With respect to protein binding, chemical profiles were retained at a lower dilution (higher concentration), but a greater dilution factor (ie, lower concentration) resulted in higher protein binding in cell medium, which in turn altered the ultimate chemical profile of bioavailable PACs. Overall, this case study demonstrates that extraction procedures, protein binding in cell culture media, and dilution factors prior to in vitro testing can all contribute to determining the final bioavailable concentrations of bioactive constituents of UVCBs in vitro. Thus, in vitro-to-in vivo extrapolation for UVCBs may require greater attention to the concentration-dependent and compound-specific differences in recovery and bioavailability.

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